Open AccessAcetylation reduces the ability of C he Y to undergo autophosphorylation
Author(s) -
Li Ru,
Chen Peng,
Gu Jing,
Deng JiaoYu
Publication year - 2013
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/1574-6968.12224
Subject(s) - autophosphorylation , acetylation , chemistry , biophysics , biochemistry , biology , phosphorylation , protein kinase a , gene
CheY , the response regulator of the chemotaxis system in E scherichia coli , can be regulated by two covalent modifications – phosphorylation and acetylation. Both covalent modifications are involved in chemotaxis, but the mechanism and role of the acetylation are still obscure. While acetylation was shown to repress the binding of CheY to its target proteins, the effect of acetylation on the ability of CheY to undergo autophosphorylate with A c P is not fully investigated. To obtain more information on the function of this acetylation, we successfully expressed and purified CheY protein with a 6 × H is‐tag on the C ‐terminus. Subsequently, acetylated CheY ( A c C he Y ) was obtained with A c C o A as the acetyl donor, and the acetylation level of A c C he Y was confirmed by Western blotting and then mass spectrometry. Using tryptophan fluorescence intensity measurements as a monitor of phosphorylation, we showed that acetylation reduces the ability of CheY to undergo autophosphorylation.